【摘要】 目的 :为探讨牛摩拉氏菌中国地方株MABL80 11限制酶的存在。方法 :采用改进的GelinasRE等方法对MABL80 11菌株的培养物分别进行超声裂解、离心、盐析、BioGel A0 .5M和磷酸纤维素P11柱层析 ,获得可切割λDNA和pBR332DNA与国际标准酶MboI切点一致的酶提取物。结果 :在改进的条件下 ,30 0 0ml培养物 ,可获得 72 0 0UMboI,纯化率为 2 3.2 % ,酶产出量高于文献报导值。结论 :表明牛摩拉氏菌中国地方株MABL80 11确含MboI限制性内切酶 ,完全可用于基因结构分析更多还原

【Abstract】 Objective: To explote the existence of restriction endonuclease in the local Moraxella bovis MABL8011. Methods: The restriction endonuclease in the cultured medium of this strain was extracted and purified with modified methods reported by Gelinas et al. The experimental procedure included sanitation, centrifugation, salt out, BioGel 0.5M, and phosphocellulose p11 column chromatography. Results: The obtained enzyme was found to be able to cut λDNA and pBR 322 DNA at the same cleavage sites as the standard enzyme Mbo I did. There was a larger amount of the enzyme isolated from the cultured medium in our experiment than that in other report. 7,200 U of Mbo I was isolated from 3,000 ml of the cultured media of MABL 8011 in the modified experiment with a purificatory rate of 23.2%. Conclusion: Our results prove that the local Moraxella bovine strain MABL 8011 can produce restriction endonuclease MboI that has biological function of cutting gene.更多还原